DFO、5-ALA不同浓度与pH值对大肠癌细胞原卟啉聚集的影响

Effects of DFO,5-ALA at Different Concentrations and pH Value on the Protoporphyrin Accumulation in Colorectal Cancer Cell

目的:探讨DFO、5-ALA不同浓度和pH值对体外诱导大肠癌细胞原卟啉Ⅸ聚积的影响,为该药物诱导大肠癌组织原卟啉Ⅸ聚积用于大肠早癌的诊断提供依据。方法:pH值分别为5.5、6.0、6.5、7.0、7.5、8;5-ALA浓度分别为0.0、0.5、1.0、1.5、2.0、2.5、3.0moml/mL;DFO浓度分别为0.0、5、10、15、20、30moml/mL的诱导培养液分别培养2小时后终止培养。高效液相色谱-荧光检测仪测定细胞内原卟啉Ⅸ含量。结果:pH值为7.0时原卟啉Ⅸ含量最高,与pH﹤6.5或pH≥7.5各组相比有显著差异(P﹤0.05);DFO浓度为10moml/mL时,5-ALA的最适诱导浓度为1.5～2.0moml/mL;5-ALA浓度为2moml/mL时,DFO的最适诱导浓度为10～20moml/mL。结论:5-ALA的最适浓度为1.5～2.0moml/mL,DFO的最适浓度为10～20moml/mL;诱导药物缓冲液的最适pH值为6.5～7。

Objective To explore the effects of DFO,5-ALA at different concentrations and pH value on the accumulation of protoporphyrin IX in colorectal carcinoma cells induced in vitro,to provide the gist for the diagnosis of early colorectal cancer on cumulation of protoporphysin IX in colorectal cancer tissue induced by durg.Methods Indu ced-culture medium were cultured for 2 hours after the termination of culture with pH values,5-ALA concentrations and DFO concentrations differ from 5.5,6.0,6.5,7.0,7.5,8;0.0,0.5,1.0,1.5,2.0,2.5,3.0 momL/mL;0.0,5,10,15,20,30 momL/mL High-performance liquid chromatography-fluorescence detection was used to measure the intracellular content of protoporphyrin Ⅸ.Results The content of protoporphyrin Ⅸ has achieved the maximus when pH=7.0,which has significant difference from which of pH <6.5 or pH≥7.5 in each group(P<0.05);the optimal induction level of 5-ALA is 1.5～2.0momL/mL with DFO level 10momL/mL;the optimal induction level of DFO is 10～20moml/mL with 5-ALA 2 momL/mL.Conclusion The optimal level of 5-ALA was 1.5～2.0 moml/mL,with DFO 10～20moml /ml.The optimal PH of experimental induction drug balanced solution is 6.5～7.